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. 1979 Jul;25(1):328–336. doi: 10.1128/iai.25.1.328-336.1979

Analysis of the effects of lipopolysaccharide on macrophages: differential phagocytic responses of C3H/HeN and C3H/HeJ macrophages in vitro.

S N Vogel, S T Marshall, D L Rosenstreich
PMCID: PMC414456  PMID: 478638

Abstract

The phenomenon of lipopolysaccharide (LPS)-induced in vitro macrophage cytotoxicity has been reported by a number of investigators but has often been difficult to reproduce and to quantitate. In this report, we have examined the effect of LPS on the ability of macrophages to ingest 51Cr-labeled, opsonized sheep erythrocytes as a method for examining the direct toxic effects of LPS on macrophages in vitro. By using this assy, we can clearly discriminate between LPS responder C3H/HeN macrophages and LPS nonresponder C3H/HeJ macrophages and demonstrate that LPS induces a profound inhibition of Fc-mediated phagocytosis in LPS responsive macrophages. Furthermore, low concentrations of LPS stimulate phagocytosis in macrophages derived for C3H/HeJ mice. The lipid A moiety of the LPS is responsible for the observed enhancement or inhibition of Fc-mediated phagocytosis. This assay was more sensitive than LPS-induced cytotoxicity, since inhibition of phagocytosis was detectable in cultures of LPS-sensitive macrophages even when cytotoxicity, assessed by trypan blue exclusion, was not. Thus, this assay represents an extremely sensitive method for analyzing the direct effects of LPS on macrophages.

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Selected References

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  1. Auletta F. J., Ausman R. M., Caldwell B. V. Development and standardization of radioimmunoassays for prostaglandins E, F, and A. Clin Chem. 1974 Dec;20(12):1580–1587. [PubMed] [Google Scholar]
  2. Berry L. J. Bacterial toxins. CRC Crit Rev Toxicol. 1977 Nov;5(3):239–318. doi: 10.3109/10408447709082601. [DOI] [PubMed] [Google Scholar]
  3. Freeman G. G. The preparation and properties of a specific polysaccharide from Bact. typhosum Ty(2): With an addendum by J. St L. Philpot, From the Department of Biochemistry, Oxford. Biochem J. 1942 Apr;36(3-4):340–356. doi: 10.1042/bj0360340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Glode L. M., Jacques A., Mergenhagen S. E., Rosenstreich D. L. Resistance of macrophages from C3H/HeJ mice to the in vitro cytotoxic effects of endotoxin. J Immunol. 1977 Jul;119(1):162–166. [PubMed] [Google Scholar]
  5. Glode L. M., Mergenhagen S. E., Rosenstreich D. L. Significant contribution of spleen cells in mediating the lethal effects of endotoxin in vivo. Infect Immun. 1976 Sep;14(3):626–630. doi: 10.1128/iai.14.3.626-630.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Glode L. M., Scher I., Osborne B., Rosenstreich D. L. Cellular mechanism of endotoxin unresponsiveness in C3H/HeJ mice. J Immunol. 1976 Feb;116(2):454–461. [PubMed] [Google Scholar]
  7. HASKINS W. T., LANDY M., MILNER K. C., RIBI E. Biological properties of parent endotoxins and lipoid fractions, with a kinetic study of acid-hydrolyzed endotoxin. J Exp Med. 1961 Nov 1;114:665–684. doi: 10.1084/jem.114.5.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. HEILMAN D. H. THE SELECTIVE TOXICITY OF ENDOTOXIN FOR PHAGOCYTIC CELLS OF THE RETICULOENDOTHELIAL SYSTEM. Int Arch Allergy Appl Immunol. 1965;26:63–79. doi: 10.1159/000229555. [DOI] [PubMed] [Google Scholar]
  9. Haas G. P., Johnson A. G., Nowotny A. Suppression of the immune response in C3H/HeJ mice by protein-free lipopolysaccharides. J Exp Med. 1978 Oct 1;148(4):1081–1086. doi: 10.1084/jem.148.4.1081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jacobs M. D., Morrison D. C. Dissociation between mitogenicity and immunogenicity of TNP-lipopolysaccharide, a T-independent antigen. J Exp Med. 1975 Jun 1;141(6):1453–1458. doi: 10.1084/jem.141.6.1453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kessel R. W., Braun W. Cytotoxicity of endotoxin in vitro. Effects on macrophages from normal guinea pigs. Aust J Exp Biol Med Sci. 1965 Aug;43(4):511–522. doi: 10.1038/icb.1965.36. [DOI] [PubMed] [Google Scholar]
  12. McIntire F. C., Sievert H. W., Barlow G. H., Finley R. A., Lee A. Y. Chemical, physical, biological properties of a lipopolysaccharide from Escherichia coli K-235. Biochemistry. 1967 Aug;6(8):2363–2372. doi: 10.1021/bi00860a011. [DOI] [PubMed] [Google Scholar]
  13. Moeller G. R., Terry L., Snyderman R. The inflammatory response and resistance to endotoxin in mice. J Immunol. 1978 Jan;120(1):116–123. [PubMed] [Google Scholar]
  14. Moore R. N., Goodrum K. J., Berry L. J. Mediation of an endotoxic effect by macrophages. J Reticuloendothel Soc. 1976 Mar;19(3):187–197. [PubMed] [Google Scholar]
  15. Morrison D. C., Betz S. J., Jacobs D. M. Isolation of a lipid A bound polypeptide responsible for "LPS-initiated" mitogenesis of C3H/HeJ spleen cells. J Exp Med. 1976 Sep 1;144(3):840–846. doi: 10.1084/jem.144.3.840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Morrison D. C., Jacobs D. M. Binding of polymyxin B to the lipid A portion of bacterial lipopolysaccharides. Immunochemistry. 1976 Oct;13(10):813–818. doi: 10.1016/0019-2791(76)90181-6. [DOI] [PubMed] [Google Scholar]
  17. Nowotny A., Behling U. H., Chang H. L. Relation of structure to function in bacterial endotoxins. VIII. Biological activities in a polysaccharide-rich fraction. J Immunol. 1975 Jul;115(1):199–203. [PubMed] [Google Scholar]
  18. Peavy D. L., Baughn R. E., Musher D. M. Strain-dependent cytotoxic effects of endotoxin for mouse peritoneal macrophages. Infect Immun. 1978 Jul;21(1):310–319. doi: 10.1128/iai.21.1.310-319.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. RIBI E., HASKINS W. T., MILNER K. C., ANACKER R. L., RITTER D. B., GOODE G., TRAPANI R. J., LANDY M. Physicochemical changes in endotoxin associated with loss of biological potency. J Bacteriol. 1962 Oct;84:803–814. doi: 10.1128/jb.84.4.803-814.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Razin E., Bauminger S., Globerson A. Effect of prostaglandins on phagocytosis of sheep erythrocytes by mouse peritoneal macrophages. J Reticuloendothel Soc. 1978 Apr;23(4):237–242. [PubMed] [Google Scholar]
  21. Rifkind D. Prevention by polymyxin B of endotoxin lethality in mice. J Bacteriol. 1967 Apr;93(4):1463–1464. doi: 10.1128/jb.93.4.1463-1464.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rosenstreich D. L., Vogel S. N., Jacques A. R., Wahl L. M., Oppenheim J. J. Macrophage sensitivity to endotoxin: genetic control by a single codominant gene. J Immunol. 1978 Nov;121(5):1664–1670. [PubMed] [Google Scholar]
  23. Rosenstreich D. L., Vogel S. N., Jacques A., Wahl L. M., Scher I., Mergenhagen S. E. Differential endotoxin sensitivity of lymphocytes and macrophages from mice with an X-linked defect in B cell maturation. J Immunol. 1978 Aug;121(2):685–690. [PubMed] [Google Scholar]
  24. Shands J. W., Jr, Peavy D. L., Gormus B. J., McGraw J. In vitro and in vivo effects of endotoxin on mouse peritoneal cells. Infect Immun. 1974 Jan;9(1):106–112. doi: 10.1128/iai.9.1.106-112.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Skidmore B. J., Chiller J. M., Weigle W. O., Riblet R., Watson J. Immunologic properties of bacterial lipopolysaccharide (LPS). III. Genetic linkage between the in vitro mitogenic and in vivo adjuvant properties of LPS. J Exp Med. 1976 Jan 1;143(1):143–150. doi: 10.1084/jem.143.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Snyderman R., Pike M. C., Fischer D. G., Koren H. S. Biologic and biochemical activities of continuous macrophage cell lines P388D1 and J774.1. J Immunol. 1977 Dec;119(6):2060–2066. [PubMed] [Google Scholar]
  27. Vogel S. N., Hansen C. T., Rosenstreich D. L. Characterization of a congenitally LPS-resistant, athymic mouse strain. J Immunol. 1979 Feb;122(2):619–622. [PubMed] [Google Scholar]
  28. Wahl L. M., Olsen C. E., Sandberg A. L., Mergenhagen S. E. Prostaglandin regulation of macrophage collagenase production. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4955–4958. doi: 10.1073/pnas.74.11.4955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wahl L. M., Rosenstreich D. L., Glode L. M., Sandberg A. L., Mergenhagen S. E. Defective prostaglandin synthesis by C3H/HeJ mouse macrophages stimulated with endotoxin preparations. Infect Immun. 1979 Jan;23(1):8–13. doi: 10.1128/iai.23.1.8-13.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

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